Department of Microbiology & Immunology, Geisel School of Medicine at Dartmouth, Hanover, NH, 03755, USA.
Genomic Research Laboratory, Service of Infectious Diseases, Geneva University Hospitals and University Medical Center, Rue Michel-Servet 1, 1211, Geneva 4, Switzerland.
Sci Rep. 2022 Sep 2;12(1):14963. doi: 10.1038/s41598-022-18815-3.
Staphylococcus aureus adapts to different environments by sensing and responding to diverse environmental cues. The responses are coordinately regulated by regulatory proteins, and small regulatory RNAs at the transcriptional and translational levels. Here, we characterized teg58, a SarA repressed sRNA, using ChIP-Seq and RNA-Seq analysis of a sarA mutant. Phenotypic and genetic analyses indicated that inactivation of teg58 led to reduced biofilm formation in a process that is independent of SarA, agr, PIA, and PSMs. RNA-Seq analysis of teg58 mutant revealed up-regulation of arginine biosynthesis genes (i.e., argGH) as well as the ability of the mutant to grow in a chemical defined medium (CDM) lacking L-arginine. Exogenous L-arginine or endogenous induction of argGH led to decreased biofilm formation in parental strains. Further analysis in vitro and in vivo demonstrated that the specific interaction between teg58 and the argGH occurred at the post-transcriptional level to repress arginine synthesis. Biochemical and genetic analyses of various arginine catabolic pathway genes demonstrated that the catabolic pathway did not play a significant role in reduced biofilm formation in the teg58 mutant. Overall, results suggest that teg58 is a regulatory sRNA that plays an important role in modulating arginine biosynthesis and biofilm formation in S. aureus.
金黄色葡萄球菌通过感知和响应不同的环境线索来适应不同的环境。这些反应在转录和翻译水平上通过调节蛋白和小调控 RNA 进行协调调节。在这里,我们使用 sarA 突变体的 ChIP-Seq 和 RNA-Seq 分析来表征 teg58,这是一种 SarA 抑制的 sRNA。表型和遗传分析表明,teg58 的失活导致生物膜形成减少,这一过程不依赖于 SarA、agr、PIA 和 PSMs。teg58 突变体的 RNA-Seq 分析显示,精氨酸生物合成基因(即 argGH)的上调以及突变体在缺乏 L-精氨酸的化学定义培养基 (CDM) 中生长的能力。外源性 L-精氨酸或 argGH 的内源性诱导导致亲本菌株生物膜形成减少。进一步的体外和体内分析表明,teg58 和 argGH 之间的特异性相互作用发生在转录后水平,以抑制精氨酸合成。对各种精氨酸分解代谢途径基因的生化和遗传分析表明,该分解代谢途径在 teg58 突变体中生物膜形成减少中没有发挥重要作用。总的来说,结果表明 teg58 是一种调节 sRNA,它在调节金黄色葡萄球菌中精氨酸生物合成和生物膜形成方面起着重要作用。